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Relationship Between Acrylamide Concentration and Enzymatic Activity in An Improved Single Fibrin Zymogram Gel System

  • Choi, Nack-Shick (Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Byoung-Young (Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Jin-Young (Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • Yoon, Kab-Seog (Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • Han, Kyoung-Yoen (Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Seung-Ho (Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology)
  • Published : 2002.03.31

Abstract

Based on the zymography analysis, Bacillus sp. DJ-4 (screened from Doen-Jang, a Korean traditional fermented food) secretes seven extracellular fibrinolytic enzymes (EFEs; 68, 64, 55, 45, 33, 27, and 13 kDa) in culture broth. These seven EFEs were analyzed by newly applied SDS-fibrin zymography combined with gradient polyacrylamide (SDS-FZGP). This improved gel system was used with a 5-20% acrylamide gradient in a fibrin zymogram gel for the separation of proteins with molecular masses from below 10kDa to over 100kDa on one gel plate. Using this system, high molecular weight bands (HMWBs) were clearly and sharply resolved. We also examined the relationship between an acrylamide concentration and the enzymatic activity of EFE using densitometric analysis.

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